RT-qPCR
One μg of total RNA was reverse-transcribed into first-strand
complementary DNA (cDNA) with Prime Script RT Reagent Kit (Perfect Real
Time, TaKaRa, Dalian, China) according to the manufacturer’s
instructions. Real-time PCR was performed with 50ng of cDNA using
One-Step SYBR PrimeScriptTM RT-PCR kit (Takara, Dalian, China). All
reactions were performed in triplicate. All primers were initially
evaluated for efficiency using relative standard curve and
electrophoresis on gel. Primer sequences are listed in Supplementary
Table 3.